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Macrophage reprogramming into a pro-healing phenotype by siRNA delivered with LBL assembled nanocomplexes for wound healing applications

Maryam Sharifiaghdam, Elnaz Shaabani Sichani, Zeynab Sharifiaghdam, Herlinde De Keersmaecker (UGent) , Bart Lucas (UGent) , Joris Lammens, Hossein Ghanbari, Ladan Teimoori-Toolabi, Chris Vervaet (UGent) , Thomas De Beer (UGent) , et al.
(2021) NANOSCALE. 13(36). p.15445-15463
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Abstract
Excessive inflammatory responses in wounds are characterized by the presence of high levels of pro-inflammatory M1 macrophages rather than pro-healing M2 macrophages, which leads to delayed wound healing. Macrophage reprogramming from the M1 to M2 phenotype through knockdown of interferon regulatory factor 5 (irf5) has emerged as a possible therapeutic strategy. While downregulation of irf5 could be achieved by siRNA, it very much depends on successful intracellular delivery by suitable siRNA carriers. Here, we report on highly stable selenium-based layer-by-layer (LBL) nanocomplexes (NCs) for siRNA delivery with polyethyleneimine (PEI-LBL-NCs) as the final polymer layer. PEI-LBL-NCs showed good protection of siRNA with only 40% siRNA release in a buffer of pH = 8.5 after 72 h or in simulated wound fluid after 4 h. PEI-LBL-NCs also proved to be able to transfect RAW 264.7 cells with irf5-siRNA, resulting in successful reprogramming to the M2 phenotype as evidenced by a 3.4 and 2.6 times decrease in NOS-2 and TNF-alpha mRNA expression levels, respectively. Moreover, irf5-siRNA transfected cells exhibited a 2.5 times increase of the healing mediator Arg-1 and a 64% increase in expression of the M2 cell surface marker CD206(+). Incubation of fibroblast cells with conditioned medium isolated from irf5-siRNA transfected RAW 264.7 cells resulted in accelerated wound healing in an in vitro scratch assay. These results show that irf5-siRNA loaded PEI-LBL-NCs are a promising therapeutic approach to tune macrophage polarization for improved wound healing.
Keywords
mRNA, retina, retinal drug delivery, innate immune system inhibition, B18R, retinal pigment epithelial cells, Müller cells, INTERFERON REGULATORY FACTOR, IN-VITRO, LIPID NANOPARTICLES, RNAI THERAPEUTICS, TISSUE-REPAIR, CHITOSAN, POLARIZATION, DNA, NANOCARRIERS, IONIZATION

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MLA
Sharifiaghdam, Maryam, et al. “Macrophage Reprogramming into a Pro-Healing Phenotype by SiRNA Delivered with LBL Assembled Nanocomplexes for Wound Healing Applications.” NANOSCALE, edited by Maryam Sharifiaghdam, vol. 13, no. 36, 2021, pp. 15445–63, doi:10.1039/d1nr03830c.
APA
Sharifiaghdam, M., Shaabani Sichani, E., Sharifiaghdam, Z., De Keersmaecker, H., Lucas, B., Lammens, J., … Fraire, J. (2021). Macrophage reprogramming into a pro-healing phenotype by siRNA delivered with LBL assembled nanocomplexes for wound healing applications. NANOSCALE, 13(36), 15445–15463. https://doi.org/10.1039/d1nr03830c
Chicago author-date
Sharifiaghdam, Maryam, Elnaz Shaabani Sichani, Zeynab Sharifiaghdam, Herlinde De Keersmaecker, Bart Lucas, Joris Lammens, Hossein Ghanbari, et al. 2021. “Macrophage Reprogramming into a Pro-Healing Phenotype by SiRNA Delivered with LBL Assembled Nanocomplexes for Wound Healing Applications.” Edited by Maryam Sharifiaghdam. NANOSCALE 13 (36): 15445–63. https://doi.org/10.1039/d1nr03830c.
Chicago author-date (all authors)
Sharifiaghdam, Maryam, Elnaz Shaabani Sichani, Zeynab Sharifiaghdam, Herlinde De Keersmaecker, Bart Lucas, Joris Lammens, Hossein Ghanbari, Ladan Teimoori-Toolabi, Chris Vervaet, Thomas De Beer, Reza Faridi-Majidi, Stefaan De Smedt, Kevin Braeckmans, and Juan Fraire. 2021. “Macrophage Reprogramming into a Pro-Healing Phenotype by SiRNA Delivered with LBL Assembled Nanocomplexes for Wound Healing Applications.” Ed by. Maryam Sharifiaghdam. NANOSCALE 13 (36): 15445–15463. doi:10.1039/d1nr03830c.
Vancouver
1.
Sharifiaghdam M, Shaabani Sichani E, Sharifiaghdam Z, De Keersmaecker H, Lucas B, Lammens J, et al. Macrophage reprogramming into a pro-healing phenotype by siRNA delivered with LBL assembled nanocomplexes for wound healing applications. Sharifiaghdam M, editor. NANOSCALE. 2021;13(36):15445–63.
IEEE
[1]
M. Sharifiaghdam et al., “Macrophage reprogramming into a pro-healing phenotype by siRNA delivered with LBL assembled nanocomplexes for wound healing applications,” NANOSCALE, vol. 13, no. 36, pp. 15445–15463, 2021.
@article{8739573,
  abstract     = {{Excessive inflammatory responses in wounds are characterized by the presence of high levels of pro-inflammatory M1 macrophages rather than pro-healing M2 macrophages, which leads to delayed wound healing. Macrophage reprogramming from the M1 to M2 phenotype through knockdown of interferon regulatory factor 5 (irf5) has emerged as a possible therapeutic strategy. While downregulation of irf5 could be achieved by siRNA, it very much depends on successful intracellular delivery by suitable siRNA carriers. Here, we report on highly stable selenium-based layer-by-layer (LBL) nanocomplexes (NCs) for siRNA delivery with polyethyleneimine (PEI-LBL-NCs) as the final polymer layer. PEI-LBL-NCs showed good protection of siRNA with only 40% siRNA release in a buffer of pH = 8.5 after 72 h or in simulated wound fluid after 4 h. PEI-LBL-NCs also proved to be able to transfect RAW 264.7 cells with irf5-siRNA, resulting in successful reprogramming to the M2 phenotype as evidenced by a 3.4 and 2.6 times decrease in NOS-2 and TNF-alpha mRNA expression levels, respectively. Moreover, irf5-siRNA transfected cells exhibited a 2.5 times increase of the healing mediator Arg-1 and a 64% increase in expression of the M2 cell surface marker CD206(+). Incubation of fibroblast cells with conditioned medium isolated from irf5-siRNA transfected RAW 264.7 cells resulted in accelerated wound healing in an in vitro scratch assay. These results show that irf5-siRNA loaded PEI-LBL-NCs are a promising therapeutic approach to tune macrophage polarization for improved wound healing.}},
  author       = {{Sharifiaghdam, Maryam and Shaabani Sichani, Elnaz and Sharifiaghdam, Zeynab and De Keersmaecker, Herlinde and Lucas, Bart and Lammens, Joris and Ghanbari, Hossein and Teimoori-Toolabi, Ladan and Vervaet, Chris and De Beer, Thomas and Faridi-Majidi, Reza and De Smedt, Stefaan and Braeckmans, Kevin and Fraire, Juan}},
  editor       = {{Sharifiaghdam, Maryam}},
  issn         = {{2040-3364}},
  journal      = {{NANOSCALE}},
  keywords     = {{mRNA,retina,retinal drug delivery,innate immune system inhibition,B18R,retinal pigment epithelial cells,Müller cells,INTERFERON REGULATORY FACTOR,IN-VITRO,LIPID NANOPARTICLES,RNAI THERAPEUTICS,TISSUE-REPAIR,CHITOSAN,POLARIZATION,DNA,NANOCARRIERS,IONIZATION}},
  language     = {{eng}},
  number       = {{36}},
  pages        = {{15445--15463}},
  title        = {{Macrophage reprogramming into a pro-healing phenotype by siRNA delivered with LBL assembled nanocomplexes for wound healing applications}},
  url          = {{http://doi.org/10.1039/d1nr03830c}},
  volume       = {{13}},
  year         = {{2021}},
}
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